Target: Dihydrofolate Reductase-Thymidilate Synthase [Leishmania Major] EC#: 1.5.1.3 NCBI Gene: 159310 Protein ID: XP_001680857.1 Organism: Leishmania Major, Strain Friedlin Etiologic Risk Group: Risk Group 2 (Parasitic Agents) Background/Disease Information: Cutaneous Leishmaniasis Essentiality: Gene/Ortholog: eco2731 (OG4_10927); Phenotype: essential; Source study: gerdes Gene/Ortholog: Tb927.7.5480 (OG4_10927); Phenotype: significant gain of fitness in procyclic forms; Source study: alsford Gene/Ortholog: Tb927.7.5480 (OG4_10927); Phenotype: significant gain of fitness in differentiation of procyclic to bloodstream forms; Source study: alsford BRENDA EC# Page:http://tinyurl.com/DHFR-TS-BRENDA NCBI Protein Page:http://tinyurl.com/L-MAJ-DHFR-TS Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): - Sigma: http://www.sigmaaldrich.com/life-science/metabolomics/enzyme-explorer/learning-center/assay-library/ec-number.html Assay Information: The DHFR assay mixture (1.0 mL) contained 25 pM Hzfolate, 0.1 mM NADPH, varying concentrations of inhibitors, 50 mM TES, 75 mM 2-mercaptoethanol, 1 mM EDTA, and 1.0 mg of BSA at pH m.0, 25 OC.” The reaction was initiated by addition of 0.5 unit of DHFR, and conversion of NADPH to NADP’ was monitored spectrophotometrically at 340 nm on a Cary-18 instrument. One unit of enzyme activity is defined as that amount of enzyme that produces 1 nmol of product/min. [3]
Cost and Quantity of Substrate Reagents and Supplier:
- Second Closest PDB Entry:3IRM - Chain Used: Chain B - Pairwise Alignment: - Query Coverage: 98% - Max Identities: 348/523 (67%) - Positives: 403/523 (77%) - Gaps: 15/523 (2%)
Current Inhibitors: 22?? Expression Information: Weak activity with folate and no activity with pterins in Leishmania Major Purification Method: Image of protein (PyMol with features delineated and shown separately):
Amino Acid Sequence: ---------------------------------------------------------------- 1 MSRAAARFKIPMPETKADFAFPSLRAFSIVVALDMQHGIGDGESIPWRVPEDMTFFKNQT 61 TLLRNKKPPTEKKRNAVVMGRKTWESVPVKFRPLKGRLNIVLSSKATVEELLAPLPEGQR 121 AAAAQDVVVVNGGLAEALRLLARPLYCSSIETAYCVGGAQVYADAMLSPCIEKLQEVYLT 181 RIYATAPACTRFFPFPPENAATAWDLASSQGRRKSEAEGLEFEICKYVPRNHEERQYLEL 241 IDRIMKTGIVKEDRTGVGTISLFGAQMRFSLRDNRLPLLTTKRVFWRGVCEELLWFLRGE 301 TSAQLLADKDIHIWDGNGSREFLDSRGLTENKEMDLGPVYGFQWRHFGADYKGFEANYDG 361 EGVDQIKLIVETIKTNPNDRRLLVTAWNPCALQKMALPPCHLLAQFYVNTDTSELSCMLY 421 QRSCDMGLGVPFNIASYALLTILIAKATGLRPGELVHTLGDAHVYRNHVDALKAQLERVP 481 HAFPTLIFKEERQYLEDYELTDMEVIDYVPHPAIKMEMAVX ---------------------------------------------------------------- Amino Acid Length of Protein: 520 Molecular Weight of Protein Using Expasy ProtParam: 58688.6 kDa Molar Extinction Coefficient at 280 nm Wavelength: 69955 or 69330 TMpred Graph:
2. Andriantsoanirina, V.; Durand, R.; Pradines, B.; Baret, E.; Bouchier, C.; Ratsimbasoa, A.; MÈnard, D., In vitro susceptibility to pyrimethamine of DHFR I164L single mutant Plasmodium falciparum. Malar J 2011, 10, 283.
3. Arrebola, R.; Olmo, A.; Reche, P.; Garvey, E. P.; Santi, D. V.; Ruiz-Perez, L. M.; Gonzalez-Pacanowska, D., Isolation and characterization of a mutant dihydrofolate reductase-thymidylate synthase from methotrexate-resistant Leishmania cells. J Biol Chem 1994, 269 (14), 10590-6.
5. Atreya, C. E.; Johnson, E. F.; Irwin, J. J.; Dow, A.; Massimine, K. M.; Coppens, I.; Stempliuk, V.; Beverley, S.; Joiner, K. A.; Shoichet, B. K.; Anderson, K. S., A molecular docking strategy identifies Eosin B as a non-active site inhibitor of protozoal bifunctional thymidylate synthase-dihydrofolate reductase. J Biol Chem 2003, 278 (16), 14092-100.
6. Bernstein, F. C.; Koetzle, T. F.; Williams, G. J.; Meyer, E. F.; Brice, M. D.; Rodgers, J. R.; Kennard, O.; Shimanouchi, T.; Tasumi, M., The Protein Data Bank: a computer-based archival file for macromolecular structures. J Mol Biol 1977, 112 (3), 535-42.
7. Beverley, S. M.; Ellenberger, T. E.; Cordingley, J. S., Primary structure of the gene encoding the bifunctional dihydrofolate reductase-thymidylate synthase of Leishmania major. Proc Natl Acad Sci U S A 1986, 83 (8), 2584-8.
8. Booth, R. G.; Selassie, C. D.; Hansch, C.; Santi, D. V., Quantitative structure-activity relationship of triazine-antifolate inhibition of Leishmania dihydrofolate reductase and cell growth. J Med Chem 1987, 30 (7), 1218-24.
9. Chitnumsub, P.; Yuvaniyama, J.; Chahomchuen, T.; Vilaivan, T.; Yuthavong, Y., Crystallization and preliminary crystallographic studies of dihydrofolate reductase-thymidylate synthase from Trypanosoma cruzi, the Chagas disease pathogen. Acta Crystallogr Sect F Struct Biol Cryst Commun 2009, 65 (Pt 11), 1175-8.
11. Ferrari, S.; Morandi, F.; Motiejunas, D.; Nerini, E.; Henrich, S.; Luciani, R.; Venturelli, A.; Lazzari, S.; CalÚ, S.; Gupta, S.; Hannaert, V.; Michels, P. A.; Wade, R. C.; Costi, M. P., Virtual screening identification of nonfolate compounds, including a CNS drug, as antiparasitic agents inhibiting pteridine reductase. J Med Chem 2011, 54 (1), 211-21.
12. Gilbert, I. H., Inhibitors of dihydrofolate reductase in Leishmania and trypanosomes. Biochim Biophys Acta 2002, 1587 (2-3), 249-57.
13. Heaslet, H.; Harris, M.; Fahnoe, K.; Sarver, R.; Putz, H.; Chang, J.; Subramanyam, C.; Barreiro, G.; Miller, J. R., Structural comparison of chromosomal and exogenous dihydrofolate reductase from Staphylococcus aureus in complex with the potent inhibitor trimethoprim. Proteins 2009, 76 (3), 706-17.
14. Hillcoat, B. L.; Nixon, P. F.; Blakley, R. L., Effect of substrate decomposition on the spectrophotometric assay of dihydrofolate reductase. Anal Biochem 1967, 21 (2), 178-89.
15. Jones, G.; Willett, P.; Glen, R. C.; Leach, A. R.; Taylor, R., Development and validation of a genetic algorithm for flexible docking. J Mol Biol 1997, 267 (3), 727-48.
16. Khabnadideh, S.; Pez, D.; Musso, A.; Brun, R.; PÈrez, L. M.; Gonz·lez-Pacanowska, D.; Gilbert, I. H., Design, synthesis and evaluation of 2,4-diaminoquinazolines as inhibitors of trypanosomal and leishmanial dihydrofolate reductase. Bioorg Med Chem 2005, 13 (7), 2637-49.
17. Knighton, D. R.; Kan, C. C.; Howland, E.; Janson, C. A.; Hostomska, Z.; Welsh, K. M.; Matthews, D. A., Structure of and kinetic channelling in bifunctional dihydrofolate reductase-thymidylate synthase. Nat Struct Biol 1994, 1 (3), 186-94.
18. Lipinski, C. A.; Lombardo, F.; Dominy, B. W.; Feeney, P. J., Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv Drug Deliv Rev 2001, 46 (1-3), 3-26.
19. Meek, T. D.; Garvey, E. P.; Santi, D. V., Purification and characterization of the bifunctional thymidylate synthetase-dihydrofolate reductase from methotrexate-resistant Leishmania tropica. Biochemistry 1985, 24 (3), 678-86.
20. Nare, B.; Hardy, L. W.; Beverley, S. M., The roles of pteridine reductase 1 and dihydrofolate reductase-thymidylate synthase in pteridine metabolism in the protozoan parasite Leishmania major. In J Biol Chem, 1997; Vol. 272, pp 13883-91.
21. Schomburg, I.; Chang, A.; Schomburg, D., BRENDA, enzyme data and metabolic information. Nucleic Acids Res 2002, 30 (1), 47-9.
EC#: 1.5.1.3
NCBI Gene: 159310
Protein ID: XP_001680857.1
Organism: Leishmania Major, Strain Friedlin
Etiologic Risk Group: Risk Group 2 (Parasitic Agents)
Background/Disease Information: Cutaneous Leishmaniasis
Essentiality:
Gene/Ortholog: eco2731 (OG4_10927); Phenotype: essential; Source study: gerdes
Gene/Ortholog: Tb927.7.5480 (OG4_10927); Phenotype: significant gain of fitness in procyclic forms; Source study: alsford
Gene/Ortholog: Tb927.7.5480 (OG4_10927); Phenotype: significant gain of fitness in differentiation of procyclic to bloodstream forms; Source study: alsford
BRENDA EC# Page: http://tinyurl.com/DHFR-TS-BRENDA
NCBI Protein Page: http://tinyurl.com/L-MAJ-DHFR-TS
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
- Sigma: http://www.sigmaaldrich.com/life-science/metabolomics/enzyme-explorer/learning-center/assay-library/ec-number.html
Assay Information:
The DHFR assay mixture (1.0 mL) contained 25 pM Hzfolate, 0.1 mM NADPH, varying concentrations of inhibitors, 50 mM TES, 75 mM 2-mercaptoethanol, 1 mM EDTA, and 1.0 mg of BSA at pH m.0, 25 OC.” The reaction was initiated by addition of 0.5 unit of DHFR, and conversion of NADPH to NADP’ was monitored spectrophotometrically at 340 nm on a Cary-18 instrument. One unit of enzyme activity is defined as that amount of enzyme that produces 1 nmol of product/min. [3]
Cost and Quantity of Substrate Reagents and Supplier:
Homology Model:
- Closest PDB Entry: 3INV
- Chain Used: Chain A
- Pairwise Alignment:
- Query Coverage: 98%
- Max Identities: 348/523 (67%)
- Positives: 403/523 (77%)
- Gaps: 15/523 (2%)
- Second Closest PDB Entry: 3IRM
- Chain Used: Chain B
- Pairwise Alignment:
- Query Coverage: 98%
- Max Identities: 348/523 (67%)
- Positives: 403/523 (77%)
- Gaps: 15/523 (2%)
Current Inhibitors: 22??
Expression Information:
Weak activity with folate and no activity with pterins in Leishmania Major
Purification Method:
Image of protein (PyMol with features delineated and shown separately):
Amino Acid Sequence:
----------------------------------------------------------------
1 MSRAAARFKIPMPETKADFAFPSLRAFSIVVALDMQHGIGDGESIPWRVPEDMTFFKNQT
61 TLLRNKKPPTEKKRNAVVMGRKTWESVPVKFRPLKGRLNIVLSSKATVEELLAPLPEGQR
121 AAAAQDVVVVNGGLAEALRLLARPLYCSSIETAYCVGGAQVYADAMLSPCIEKLQEVYLT
181 RIYATAPACTRFFPFPPENAATAWDLASSQGRRKSEAEGLEFEICKYVPRNHEERQYLEL
241 IDRIMKTGIVKEDRTGVGTISLFGAQMRFSLRDNRLPLLTTKRVFWRGVCEELLWFLRGE
301 TSAQLLADKDIHIWDGNGSREFLDSRGLTENKEMDLGPVYGFQWRHFGADYKGFEANYDG
361 EGVDQIKLIVETIKTNPNDRRLLVTAWNPCALQKMALPPCHLLAQFYVNTDTSELSCMLY
421 QRSCDMGLGVPFNIASYALLTILIAKATGLRPGELVHTLGDAHVYRNHVDALKAQLERVP
481 HAFPTLIFKEERQYLEDYELTDMEVIDYVPHPAIKMEMAVX
----------------------------------------------------------------
Amino Acid Length of Protein: 520
Molecular Weight of Protein Using Expasy ProtParam: 58688.6 kDa
Molar Extinction Coefficient at 280 nm Wavelength: 69955 or 69330
TMpred Graph:
CDS Gene Sequence:
----------------------------------------------------------------
1 ATGTCCAGGGCAGCTGCGAGGTTTAAGATTCCGATGCCGGAGACGAAGGCAGACTTTGCT
61 TTCCCCTCCCTGCGCGCCTTCTCCATCGTCGTGGCCCTCGATATGCAGCACGGCATCGGC
121 GACGGCGAGTCGATCCCGTGGCGGGTGCCGGAGGACATGACGTTCTTCAAGAACCAGACG
181 ACGCTGCTGCGCAACAAGAAGCCGCCGACGGAGAAGAAGCGCAACGCCGTCGTGATGGGC
241 CGCAAGACTTGGGAGAGCGTCCCGGTAAAGTTCCGACCACTCAAGGGACGGCTGAACATC
301 GTGTTATCCTCGAAGGCCACCGTCGAGGAGCTTCTGGCGCCGCTGCCGGAGGGACAGCGC
361 GCGGCGGCGGCGCAGGATGTGGTGGTGGTGAACGGCGGTCTGGCCGAGGCGCTCCGCCTC
421 CTCGCACGCCCGCTGTACTGCAGCTCCATCGAGACAGCGTATTGCGTCGGTGGTGCGCAG
481 GTTTACGCGGACGCCATGCTGTCGCCGTGCATCGAGAAACTGCAGGAAGTGTACCTGACC
541 CGCATCTACGCGACGGCGCCTGCGTGTACGCGCTTCTTTCCGTTTCCGCCCGAGAACGCG
601 GCCACGGCGTGGGACCTGGCGTCGTCTCAGGGACGCCGCAAGAGCGAGGCGGAGGGCCTC
661 GAGTTCGAGATCTGCAAGTACGTGCCGCGCAACCACGAGGAGCGGCAGTACCTTGAGCTG
721 ATTGACCGCATCATGAAGACGGGGATCGTGAAGGAGGACCGCACCGGCGTGGGCACCATC
781 AGCCTCTTCGGCGCCCAGATGCGCTTCTCCCTACGCGACAACCGCCTGCCGCTGCTGACG
841 ACGAAGCGTGTCTTCTGGCGCGGCGTGTGCGAGGAGCTGCTGTGGTTCCTGCGCGGGGAG
901 ACGAGTGCGCAGCTGCTGGCAGACAAGGACATTCACATCTGGGACGGCAACGGTTCGCGC
961 GAGTTTCTCGACAGCCGCGGCTTGACAGAGAATAAGGAGATGGACCTCGGCCCTGTCTAC
1021 GGCTTCCAGTGGCGCCACTTCGGGGCAGATTACAAGGGGTTTGAAGCGAACTACGACGGC
1081 GAAGGGGTGGACCAGATCAAGCTCATCGTGGAGACCATCAAGACGAACCCGAACGACCGC
1141 CGCCTCCTAGTCACTGCCTGGAACCCGTGCGCGCTGCAAAAGATGGCGCTGCCGCCGTGC
1201 CACTTGCTTGCTCAGTTCTACGTGAACACAGACACGAGCGAGCTATCCTGCATGTTGTAC
1261 CAGCGCTCGTGTGACATGGGTCTTGGCGTCCCCTTCAACATTGCCTCCTACGCGCTGCTC
1321 ACCATCCTCATTGCCAAGGCGACGGGTCTGCGGCCTGGTGAGCTTGTGCACACCCTCGGC
1381 GACGCCCACGTCTACCGCAACCACGTTGATGCCCTCAAGGCGCAGCTCGAGCGAGTCCCG
1441 CACGCGTTCCCGACCCTCATCTTCAAGGAGGAGCGGCAGTACCTCGAGGACTACGAGTTG
1501 ACGGACATGGAGGTGATCGACTACGTTCCACACCCGGCGATCAAGATGGAGATGGCCGTA
1561 TAG
----------------------------------------------------------------
GC% Content: 63.08%
Codon Optimized CDS Gene Sequence:
----------------------------------------------------------------
1 ATGTCTCGTGCCGCAGCCCGCTTTAAGATCCCGATGCCTGAGACCAAAGCGGACTTCGCG
61 TTCCCGTCCCTGCGTGCTTTCTCTATTGTAGTAGCGCTGGACATGCAGCACGGTATTGGT
121 GACGGCGAGTCTATTCCGTGGCGTGTACCGGAGGATATGACCTTTTTCAAGAATCAGACC
181 ACTCTGCTGCGCAACAAAAAACCGCCGACTGAGAAAAAACGCAATGCGGTTGTTATGGGC
241 CGCAAAACTTGGGAATCTGTCCCTGTTAAATTCCGTCCACTCAAAGGTCGTCTGAACATC
301 GTTCTGTCTTCTAAAGCGACTGTTGAGGAACTGCTCGCACCTCTCCCAGAAGGCCAGCGC
361 GCAGCGGCAGCACAAGATGTTGTTGTGGTTAACGGTGGCCTGGCGGAAGCCCTCCGCCTC
421 CTCGCGCGTCCGCTCTACTGTAGCTCTATCGAGACCGCCTACTGCGTAGGTGGTGCACAG
481 GTTTACGCAGATGCCATGCTCTCCCCGTGCATCGAGAAGCTGCAGGAAGTATACCTGACT
541 CGCATTTACGCGACGGCTCCGGCGTGCACTCGTTTCTTCCCGTTCCCTCCGGAAAACGCG
601 GCGACTGCGTGGGATCTCGCGTCCTCTCAAGGTCGCCGTAAATCTGAAGCGGAAGGCCTG
661 GAATTTGAAATCTGCAAATACGTTCCGCGTAATCACGAAGAGCGTCAGTATCTCGAACTC
721 ATCGACCGTATCATGAAAACTGGCATCGTGAAAGAAGACCGTACCGGTGTTGGCACCATC
781 AGCCTGTTTGGTGCTCAGATGCGTTTCTCTCTGCGTGACAACCGTCTGCCACTGCTGACT
841 ACGAAACGTGTGTTCTGGCGTGGTGTTTGTGAGGAGCTGCTGTGGTTCCTCCGCGGTGAG
901 ACGTCCGCGCAACTCCTGGCCGACAAAGACATTCACATCTGGGACGGCAATGGTTCCCGC
961 GAGTTCCTGGACTCTCGTGGCCTCACCGAGAACAAGGAGATGGACCTGGGTCCGGTGTAC
1021 GGCTTTCAGTGGCGTCACTTCGGTGCGGACTACAAAGGTTTCGAAGCGAACTACGATGGC
1081 GAAGGTGTTGACCAGATCAAACTGATCGTTGAAACCATCAAAACCAACCCTAACGACCGT
1141 CGTCTCCTGGTAACCGCGTGGAACCCGTGCGCCCTGCAAAAGATGGCGCTCCCGCCGTGT
1201 CATCTGCTGGCGCAATTCTACGTGAACACCGACACCTCTGAACTGTCTTGCATGCTGTAC
1261 CAGCGTTCTTGCGACATGGGCCTGGGCGTTCCTTTCAACATCGCGTCTTACGCGCTCCTG
1321 ACCATTCTCATCGCGAAAGCCACGGGCCTGCGTCCGGGCGAACTGGTCCATACCCTGGGT
1381 GACGCGCATGTTTACCGCAACCACGTTGATGCGCTGAAGGCCCAGCTCGAACGTGTTCCA
1441 CATGCATTCCCGACCCTGATTTTCAAAGAGGAACGTCAATACCTGGAGGACTATGAACTG
1501 ACTGATATGGAAGTGATCGATTATGTACCACATCCGGCGATTAAGATGGAAATGGCTGTC
1561 TAA
----------------------------------------------------------------
GC% Content: 55.21%
Virtual Screening Work
Non-Prepped Control Library:
Library Contains the Following Ligands : KKT, MGH, 1WL, AHM, IDK, ASP, DHF, P65, MMV, WRA, DQ1, 2CY, TMQ, MTX, C50, UMP, and 1CY
Undergraduate Research Forum Poster:
Relevant Literature, References:
1. Parasites†-†Leishmaniasis. http://www.cdc.gov/parasites/leishmaniasis/ (accessed October 12).
2. Andriantsoanirina, V.; Durand, R.; Pradines, B.; Baret, E.; Bouchier, C.; Ratsimbasoa, A.; MÈnard, D., In vitro susceptibility to pyrimethamine of DHFR I164L single mutant Plasmodium falciparum. Malar J 2011, 10, 283.
3. Arrebola, R.; Olmo, A.; Reche, P.; Garvey, E. P.; Santi, D. V.; Ruiz-Perez, L. M.; Gonzalez-Pacanowska, D., Isolation and characterization of a mutant dihydrofolate reductase-thymidylate synthase from methotrexate-resistant Leishmania cells. J Biol Chem 1994, 269 (14), 10590-6.
4. Artimo, P.; Jonnalagedda, M.; Arnold, K.; Baratin, D.; Csardi, G.; de Castro, E.; Duvaud, S.; Flegel, V.; Fortier, A.; Gasteiger, E.; Grosdidier, A.; Hernandez, C.; Ioannidis, V.; Kuznetsov, D.; Liechti, R.; Moretti, S.; Mostaguir, K.; Redaschi, N.; Rossier, G.; Xenarios, I.; Stockinger, H., ExPASy: SIB bioinformatics resource portal. Nucleic Acids Res 2012, 40 (Web Server issue), W597-603.
5. Atreya, C. E.; Johnson, E. F.; Irwin, J. J.; Dow, A.; Massimine, K. M.; Coppens, I.; Stempliuk, V.; Beverley, S.; Joiner, K. A.; Shoichet, B. K.; Anderson, K. S., A molecular docking strategy identifies Eosin B as a non-active site inhibitor of protozoal bifunctional thymidylate synthase-dihydrofolate reductase. J Biol Chem 2003, 278 (16), 14092-100.
6. Bernstein, F. C.; Koetzle, T. F.; Williams, G. J.; Meyer, E. F.; Brice, M. D.; Rodgers, J. R.; Kennard, O.; Shimanouchi, T.; Tasumi, M., The Protein Data Bank: a computer-based archival file for macromolecular structures. J Mol Biol 1977, 112 (3), 535-42.
7. Beverley, S. M.; Ellenberger, T. E.; Cordingley, J. S., Primary structure of the gene encoding the bifunctional dihydrofolate reductase-thymidylate synthase of Leishmania major. Proc Natl Acad Sci U S A 1986, 83 (8), 2584-8.
8. Booth, R. G.; Selassie, C. D.; Hansch, C.; Santi, D. V., Quantitative structure-activity relationship of triazine-antifolate inhibition of Leishmania dihydrofolate reductase and cell growth. J Med Chem 1987, 30 (7), 1218-24.
9. Chitnumsub, P.; Yuvaniyama, J.; Chahomchuen, T.; Vilaivan, T.; Yuthavong, Y., Crystallization and preliminary crystallographic studies of dihydrofolate reductase-thymidylate synthase from Trypanosoma cruzi, the Chagas disease pathogen. Acta Crystallogr Sect F Struct Biol Cryst Commun 2009, 65 (Pt 11), 1175-8.
10. Chitnumsub, P.; Yuvaniyama, J.; Vilaivan, T.; Vanichtanankul, J.; Kamchonwongpaisan, S.; Yuthavong, Y., Structural basis of antifolate inhibition of Trypanosoma cruzi Dihydrofolate Reductase-Thymidylate Synthase. 2011.
11. Ferrari, S.; Morandi, F.; Motiejunas, D.; Nerini, E.; Henrich, S.; Luciani, R.; Venturelli, A.; Lazzari, S.; CalÚ, S.; Gupta, S.; Hannaert, V.; Michels, P. A.; Wade, R. C.; Costi, M. P., Virtual screening identification of nonfolate compounds, including a CNS drug, as antiparasitic agents inhibiting pteridine reductase. J Med Chem 2011, 54 (1), 211-21.
12. Gilbert, I. H., Inhibitors of dihydrofolate reductase in Leishmania and trypanosomes. Biochim Biophys Acta 2002, 1587 (2-3), 249-57.
13. Heaslet, H.; Harris, M.; Fahnoe, K.; Sarver, R.; Putz, H.; Chang, J.; Subramanyam, C.; Barreiro, G.; Miller, J. R., Structural comparison of chromosomal and exogenous dihydrofolate reductase from Staphylococcus aureus in complex with the potent inhibitor trimethoprim. Proteins 2009, 76 (3), 706-17.
14. Hillcoat, B. L.; Nixon, P. F.; Blakley, R. L., Effect of substrate decomposition on the spectrophotometric assay of dihydrofolate reductase. Anal Biochem 1967, 21 (2), 178-89.
15. Jones, G.; Willett, P.; Glen, R. C.; Leach, A. R.; Taylor, R., Development and validation of a genetic algorithm for flexible docking. J Mol Biol 1997, 267 (3), 727-48.
16. Khabnadideh, S.; Pez, D.; Musso, A.; Brun, R.; PÈrez, L. M.; Gonz·lez-Pacanowska, D.; Gilbert, I. H., Design, synthesis and evaluation of 2,4-diaminoquinazolines as inhibitors of trypanosomal and leishmanial dihydrofolate reductase. Bioorg Med Chem 2005, 13 (7), 2637-49.
17. Knighton, D. R.; Kan, C. C.; Howland, E.; Janson, C. A.; Hostomska, Z.; Welsh, K. M.; Matthews, D. A., Structure of and kinetic channelling in bifunctional dihydrofolate reductase-thymidylate synthase. Nat Struct Biol 1994, 1 (3), 186-94.
18. Lipinski, C. A.; Lombardo, F.; Dominy, B. W.; Feeney, P. J., Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv Drug Deliv Rev 2001, 46 (1-3), 3-26.
19. Meek, T. D.; Garvey, E. P.; Santi, D. V., Purification and characterization of the bifunctional thymidylate synthetase-dihydrofolate reductase from methotrexate-resistant Leishmania tropica. Biochemistry 1985, 24 (3), 678-86.
20. Nare, B.; Hardy, L. W.; Beverley, S. M., The roles of pteridine reductase 1 and dihydrofolate reductase-thymidylate synthase in pteridine metabolism in the protozoan parasite Leishmania major. In J Biol Chem, 1997; Vol. 272, pp 13883-91.
21. Schomburg, I.; Chang, A.; Schomburg, D., BRENDA, enzyme data and metabolic information. Nucleic Acids Res 2002, 30 (1), 47-9.